• Environmental Analysis Health and Toxicology
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• v.31
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• pp.10.1-10.8
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• 2016

Objectives Aromatase inhibitors that block estrogen synthesis are a proven first-line hormonal therapy for postmenopausal breast cancer. Although it is known that standardized extract of Ginkgo biloba (EGb761) induces anti-carcinogenic effects like the aromatase inhibitors, the effects of EGb761 on steroidogenesis have not been studied yet. Therefore, the effects of EGb761 on steroidogenesis and aromatase activity was studied using a H295R cell model, which was a good in vitro model to predict effects on human adrenal steroidogenesis. Methods Cortisol, aldosterone, testosterone, and $17{\beta}$-estradiol were evaluated in the H295R cells by competitive enzyme-linked immunospecific assay after exposure to EGb761. Real-time polymerase chain reaction were performed to evaluate effects on critical genes in steroid hormone production, specifically cytochrome P450 (CYP11/ 17/19/21) and the hydroxysteroid dehydrogenases ($3{\beta}$-HSD2 and $17{\beta}$-HSD1/4). Finally, aromatase activities were measured with a tritiated water-release assay and by western blotting analysis. Results H295R cells exposed to EGb761 (10 and $100{\mu}g/mL$) showed a significant decrease in $17{\beta}$-estradiol and testosterone, but no change in aldosterone or cortisol. Genes (CYP19 and $17{\beta}$-HSD1) related to the estrogen steroidogenesis were significantly decreased by EGb761. EGb761 treatment of H295R cells resulted in a significant decrease of aromatase activity as measured by the direct and indirect assays. The coding sequence/Exon PII of CYP19 gene transcript and protein level of CYP19 were significantly decreased by EGb761. Conclusions These results suggest that EGb761 could regulate steroidogenesis-related genes such as CYP19 and $17{\beta}$-HSD1, and lead to a decrease in $17{\beta}$-estradiol and testosterone. The present study provides good information on potential therapeutic effects of EGb761 on estrogen dependent breast cancer.

• The Korean Journal of Physiology and Pharmacology
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• v.21 no.2
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• pp.259-265
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• 2017

Excessive influx and the subsequent rapid cytosolic elevation of $Ca^{2+}$ in neurons is the major cause to induce hyperexcitability and irreversible cell damage although it is an essential ion for cellular signalings. Therefore, most neurons exhibit several cellular mechanisms to homeostatically regulate cytosolic $Ca^{2+}$ level in normal as well as pathological conditions. Delayed rectifier $K^+$ channels ($I_{DR}$ channels) play a role to suppress membrane excitability by inducing $K^+$ outflow in various conditions, indicating their potential role in preventing pathogenic conditions and cell damage under $Ca^{2+}$-mediated excitotoxic conditions. In the present study, we electrophysiologically evaluated the response of $I_{DR}$ channels to hyperexcitable conditions induced by high $Ca^{2+}$ pretreatment (3.6 mM, for 24 hours) in cultured hippocampal neurons. In results, high $Ca^{2+}$-treatment significantly increased the amplitude of $I_{DR}$ without changes of gating kinetics. Nimodipine but not APV blocked $Ca^{2+}$-induced $I_{DR}$ enhancement, confirming that the change of $I_{DR}$ might be targeted by $Ca^{2+}$ influx through voltage-dependent $Ca^{2+}$ channels (VDCCs) rather than NMDA receptors (NMDARs). The VDCC-mediated $I_{DR}$ enhancement was not affected by either $Ca^{2+}$-induced $Ca^{2+}$ release (CICR) or small conductance $Ca^{2+}$-activated $K^+$ channels (SK channels). Furthermore, PP2 but not H89 completely abolished $I_{DR}$ enhancement under high $Ca^{2+}$ condition, indicating that the activation of Src family tyrosine kinases (SFKs) is required for $Ca^{2+}$-mediated $I_{DR}$ enhancement. Thus, SFKs may be sensitive to excessive $Ca^{2+}$ influx through VDCCs and enhance $I_{DR}$ to activate a neuroprotective mechanism against $Ca^{2+}$-mediated hyperexcitability in neurons.

• Korean Journal of Clinical Laboratory Science
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• v.49 no.2
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• pp.99-107
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• 2017

A species of Cordyceps, an ingredient in Chinese traditional medicine well-known for its major component, cordycepin (3'-deoxyadenosine), has been known to have antiplatelet effects; however, its effects on regulation of phosphoprotein have not been fully elucidated. In this study, we investigated how cordycepin regulates the phosphoprotein, including phosphatidylinositol 3-kinase (PI3K)/Akt and p38, to inhibit platelet aggregation, which are concerned with fibrinogen binding to glycoprotein IIb/IIIa (${\alpha}IIb/{\beta}_3$) and granule secretion in platelets. Our finding suggests that cordycepin inhibits collagen-induced platelet aggregation with $261.1{\mu}M$ of $IC_{50}$ and also inhibits fibrinogen binding to ${\alpha}IIb/{\beta}_3$ by a suppression of PI3K/Akt phosphorylation in a dose dependent manner. In addition, cordycepin further showed to inhibit collagen-induced p38 phosphorylation, reducing granule secretion (i.e. ATP- and serotonin-release) and thromboxane $A_2$ ($TXA_2$) production without regulating cyclooxygenase-1 (COX-1) and thromboxane A synthase (TXAS) activities, as well as phospholipase $C-{\gamma}_2$ ($PLC-{\gamma}_2$) phosphorylation. In conclusion, these results demonstrate that cordycepin-mediated antiplatelet effects were due to the inhibition of fibrinogen binding to ${\alpha}IIb/{\beta}_3$ via the suppression of PI3K/Akt phosphorylation and inhibition of granule secretion & $TXA_2$ production by suppressing p38 phosphorylation. These results strongly indicate that cordycepin might have therapeutic or preventive potential for platelet aggregation-mediated disorders, regulating the phosphoprotein, including PI3K/Akt and p38.

• Journal of Plant Biotechnology
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• v.35 no.4
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• pp.291-298
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• 2008

Methylmercury, an organic derivative, is the principal form of mercury that biomagnifies and causes neurodegenerative symptoms in animals. In recent years, living modified organism (LMO) resulting from biotechnology has played a highly visible and controversial role. Despite the potential benefits of this technology, public concerns have been raised about the environmental risk of LMO. The concern on the risk from LMO release has urged efforts to evaluate and manage the risks of the LMO. To build up the capacity building of risk assessment method for LMO used environmental remediation, we engineered Solanum nigrum L, expressing the modified bacterial gene, merB, encoding organomercurial lyase. Two independently isolated transgenic lines produced merB RNA. Transgenic Solanum nigrum leaf discs expressing merB gene showed organic mercury resistance, forming shoots well on growth medium containing $0.5{\mu}M$ methylmercury (II) chloride and $1{\mu}M$ phenylmercuric acetate while control plants breached. Transgenic merB seeds germinated and grew on growth medium containing $2{\mu}M$ methylmercury (II) chloride and phenylmercuric acetate. The merB transgenic plants will be used for risk assessment of natural environment.

• Journal of the Korean Chemical Society
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• v.50 no.6
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• pp.464-470
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• 2006

the coricosteroid drug dexamethasone is an efficacious antiinflammatory drug, it is difficult to formulate in an injectable formulation due to its poor aqueous solubility. A lipid-based nanosphere formulation containing dexamethasone was designed for solubilization of the drug in aqueous solution and sustained release of the drug from the nanosphere. The lipid nanospheres, composed of phospholipid, cholesterol and cationic lipid, were prepared by self emulsification-solvent diffusion method followed by diafiltration. Physicochemical characteristics such as mean particle diameter, zeta potential and drug loading efficiency of the lipid nanospheres were investigated according to the variation of either the kind of lipid or the lipid composition. The lipid nanospheres had a mean diameter 80-120 nm and dexamethasone loading efficiency of greater than 80%. The drug loading efficiency increased with the increase of the length of aliphatic chain attached to the phospholipid. However, the drug loading efficiency was inversely proportional to the increase of cholesterol content in the lipid composition. The lipid nanosphere could not be prepared without the use of cationic lipid and the drug loading efficiency was proportional to the increase of cationic lipid content. The lipid nanospheres containing dexamethasone are a promising novel drug carrier for an injectable formulation of the poorly water-soluble drugs.

• Applied Biological Chemistry
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• v.50 no.3
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• pp.228-232
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• 2007

This study was performed to investigate the cytotoxic activity from Vitex rotundifolia. V. rotundifolia was extracted with methanol, ethanol, and acetone, and then the cytotoxic effect of these extracts was measured by the MTT reduction assay and morphological assay on the HT-29 human colon carcinoma cells. Among the three extracts, the acetone extract showed the highest cytotoxic activity on the HT-29 cells in a dose-dependent manner with an $IC_{50}$ value of 10 ${\mu}g/ml$. The acetone extract was further fractionated sequentially with n-hexane, diethyl ether, ethyl acetate, and water layer according to the degree of polarity. The n-hexane layer among the fractioned layers showed inhibitory activity on the growth of HT-29 cells. In addition, n-hexane layer also showed the cytotoxic activity against SW620 human colon carcinoma cells. These result indicated that extracts of V. rotundifolia may contain bioactive materials and could be potential candidates as chemotherapeutic agents against human colon carcinoma cells.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.2
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• pp.192-197
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• 2017

Objective: The study was conducted to evaluate the stability of commercial coated lipase (CT-LIP) in vitro. Methods: The capsules were tested under different conditions with a range of temperature, pH, dry heat treatment and steaming treatment, simulated gastric fluid (SGF) and simulated intestinal fluid (SIF) in this work, respectively. Free lipase (uncoated lipase, UC-LIP) was the control group. Lipase relative activities measured in various treatments were used as a reference frame to characterize the stability. Results: The lipase activities were decreased with increasing temperatures (p<0.05), and there was a markedly decline (p<0.01) in lipase comparative activities of UC-LIP at $80^{\circ}C$ compared with CT-LIP group. Higher relative activities of lipase were observed in CT-LIP group compared with the free one under acidic ambient (pH 3 to 7) and an alkaline medium (pH 8 to 12). Residual lipase activities of CT-LIP group were increased (p<0.05) by 5.67% and 35.60% in dry heat and hydrothermal treatments, respectively. The lipase relative activity profile of CT-LIP was raised at first and dropped subsequently (p<0.05) compared with constantly reduced tendency of UC-LIP exposed to both SGF and SIF. Conclusion: The results suggest that the CT-LIP possesses relatively higher stability in comparison with the UC-LIP in vitro. The CT-LIP could retain the potential property to provide sustained release of lipase and thus improved its bioavailability in the gastrointestinal tract.

• The Korean Journal of Food And Nutrition
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• v.30 no.2
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• pp.211-217
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• 2017

In the present study, we investigated the protective effect of various grain methanolic extracts against UVB-induced photo-aging in human skin fibroblasts. Various grain methanolic extracts were evaluated for their antioxidant compounds and activities. 2,2-Ddiphenyl-1-picryhydrazyl radical (DPPH) and ABTS 2,2-azino-bris-(3-ethylbenzoth iazoline-6-sulphonic acid) radical cation scavenging activities have been used to measure the relative antioxidant activities of extracts from grains. The content of total polyphenolics in the extracts were evaluated using spectrophotometric methods. Human skin fibroblast (Hs68) cells were pretreated with various grain methanolic extracts ($25{\mu}g/mL$). Skin toxicity was simulated by exposing the cells to UVB ($30mJ/cm^2$) irradiation. In response to the UVB-irradiation, an increased amount of matrix metallo-proteinases (MMPs) release was observed, whereas pretreatment of various grain methanolic extracts significantly inhibited the production of MMP-1 in Hs68 cells. We also found that pretreatment of the extracts significantly decreased UVB-induced reactive oxygen species and significantly increased total collagen content in Hs68 cells. These results provide that grains could be regarded as a potential ingredient in natural cosmetics, used for UVB protection.

• BMB Reports
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• v.37 no.3
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• pp.282-291
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• 2004

Phytases catalyze the release of phosphate from phytic acid. Phytase-producing microorganisms were selected by culturing the soil extracts on agar plates containing phytic acid. Two hundred colonies that exhibited potential phytase activity were selected for further study. The colony showing the highest phytase activity was identified as Aspergillus niger and designated strain 113. The phytase gene from A. niger 113 (phyI1) was isolated, cloned, and characterized. The nucleotide and deduced amino acid sequence identity between phyI1 and phyA from NRRL3135 were 90% and 98%, respectively. The identity between phyI1 and phyA from SK-57 was 89% and 96%. A synthetic phytase gene, phyI1s, was synthesized by successive PCR and transformed into the yeast expression vector carrying a signal peptide that was designed and synthesized using P. pastoris biased codon. For the phytase expression and secretion, the construct was integrated into the genome of P. pastoris by homologous recombination. Over-expressing strains were selected and fermented. It was discovered that ~4.2 g phytase could be purified from one liter of culture fluid. The activity of the resulting phytase was 9.5 U/mg. Due to the heavy glycosylation, the expressed phytase varied in size (120, 95, 85, and 64 kDa), but could be deglycosylated to a homogeneous 64 kDa species. An enzymatic kinetics analysis showed that the phytase had two pH optima (pH 2.0 and pH 5.0) and an optimum temperature of $60^{\circ}C$.

• Journal of Life Science
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• v.21 no.5
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• pp.678-683
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• 2011

The objective of this study was to evaluate the skin inflammation effects of three herb mixture extracts, Ligularia fischeri, Solidago virga-aurea and Aruncus dioicus, which are from Ullung island in Korea. Regulatory mechanisms of cytokines and nitric oxide (NO) are involved in the immunological activity of Raw 264.7 cells. Tested cells were pretreated with 70% acetone extracts of Ligularia fischeri, Solidago virga-aurea and Aruncus dioicus (LSA-A) and further cultured for an appropriated time after lipopolyssacharide (LPS) addition. During the entire experimental period, 1, 10, and 100 ${\mu}g/ml$ of LSA-A had no cytotoxicity. In these concentrations, LSA-A inhibited the production of NO and prostaglandin $E_2$ ($PGE_2$), tumor necorsis factor-a (TNF-a), interleukin-1${\beta}$ (IL-1${\beta}$), interleukin-6 (IL-6) expression of inducible NO synthase (iNOS), and cyclooxygenase-2 (COX-2). LSA-A showed a 60% $PGE_2$ inhibition rate at 100 ${\mu}g/ml$. iNOS and COX-2 inhibition activities were 54%, and 65% at 100 ${\mu}g/ml$, respectively. In addition, LSA-A extract reduced the release of inflammatory cytokines including TNF-a, IL-1${\beta}$ and IL-6. These results suggest that LSA-A may have significant effects on inflammatory factors, and may be a potential anti-inflammatory therapeutic agent.